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Chloroplast

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Chloroplast and Mitochondrion in Plant Cells

Introduction:
Mitochondrion and chloroplast are two organelles that are very important to organisms. Both provide energy and nourishment to the cell. While chloroplast is found in plant cells only, mitochondrion is found in both animal and plant cells. It is believed that these two organelles were once bacterial cells on their own, but then they were engulfed by other bacteria. This theory is called the Endosymbiotic Theory. This theory is proven by the fact that chloroplast and mitochondrion are the only organelles, within a cell, that have their own DNA. They also use this DNA to produce their own enzymes and proteins. Further proof is shown by the fact that both these organelles have a double layer of membrane surrounding them, and that they replicate as a bacteria does.
Purpose:
* Prepare a supernatant of spinach leaf * Demonstrate proper techniques involved in using centrifuge * Using filtration and centrifugation, separate the chloroplast from other organelles in a spinach leaf * Identify mitochondrion in an onion cell
Materials:
* Fresh spinach leaves * Grinding solution * 0.33 M sorbitol * 10mM sodium pyrophosphate (NaPO) * 4mMMgCl * 2mM Ascorbic Acid * Adjust pH to 6.5 with HCl * Chopping board and knife * Chilled mortar and pestle * Cheesecloth * Refrigerated preparative centrifuge * Suspension solution * 0.33 M Sorbitol * 2mMEDTA * 1rnMMgCl * 50rnMHEPES * Adjust pH to 7.6 with NaOH * Hematocytometer and microscope * Ice bath * Sprig of elodea
Procedures:
Part 1: Choroplasts from Spinach Leaves 1. Prepare an ice bath and pre-cool all glassware to be used. 2. Select several fresh spinach leaves and remove the large veins by tearing them loose from the leaves. Weigh out 4.0 grams of

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